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Detection of CD133 (prominin-1) in a human hepatoblastoma cell line (HuH-6 clone 5).

Akita M, Tanaka K, Murai N, Matsumoto S, Fujita K, Takaki T, Nishiyama H - Microsc. Res. Tech. (2013)

Bottom Line: An antibody against CD133 decreased cell migration.Methyl-β-cyclodextrin treatment decreased cell adhesion as well as lamellipodium and filopodium formation.The results suggest that CD133 membrane localization plays a role in tumor cell adhesion and migration.

View Article: PubMed Central - PubMed

Affiliation: Division of Morphological Science, Biomedical Research Center, Saitama Medical University, Iruma-gun, Saitama, Japan. makita@saitama-med.ac.jp

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Related in: MedlinePlus

Localization of CD133 using phase contrast microscopy and ASEM. a: Phase contrast micrograph showing a lamellipodium (arrowhead) and filopodia (arrows). Scale bar, 40 µm. b, c: Localization of CD133 by using ASEM. Cell-surface CD133 was labeled with nanogold, followed by gold enhancement. The clustered particles form a white spot approximately 50 nm in size, mainly in the lamellipodium and filopodia. Scale bar, 1 µm. b: Enlarged ASEM image of the lamellipodium indicated by an arrowhead in Figure 4a. c: Enlarged ASEM image of the filopodia indicated by arrows in Figure 4a.
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fig04: Localization of CD133 using phase contrast microscopy and ASEM. a: Phase contrast micrograph showing a lamellipodium (arrowhead) and filopodia (arrows). Scale bar, 40 µm. b, c: Localization of CD133 by using ASEM. Cell-surface CD133 was labeled with nanogold, followed by gold enhancement. The clustered particles form a white spot approximately 50 nm in size, mainly in the lamellipodium and filopodia. Scale bar, 1 µm. b: Enlarged ASEM image of the lamellipodium indicated by an arrowhead in Figure 4a. c: Enlarged ASEM image of the filopodia indicated by arrows in Figure 4a.

Mentions: Under a light microscope, strong positive results were seen for CD133 at the cell periphery (Fig. 1), where it co-localized with F-actin (Figs. 2a–2c). Preliminary examination using ASEM also showed that CD133 was localized in membrane ruffles, which occur at the peripheral regions of the cell (Fig. 3). The ASEM enables direct observation of subcellular structures, unlike phase contrast microscopy. Lamellipodia, characteristic features at the leading edge of the cell, were observed. Filopodia were observed as slender cytoplasmic projections that extended beyond the leading edge of lamellipodia. We labeled cell-surface CD133 with nanogold, followed by gold enhancement. The clustered particles formed a white spot, up to 50 nm in size, mainly in the membrane ruffles formed by lamellipodia and filopodia (Fig. 4). Counterstaining with phosphotungstic acid revealed the details of the cells, and ASEM detected details of the ventral cell surface (Fig. 5). When the Aclar® plastic film was set upside down, the dorsal surface of the cell could be observed. CD133 was mainly localized in microvillus-like plasma membrane protrusions (microspikes) on the dorsal surface of the cells Figs.(6 and 7).


Detection of CD133 (prominin-1) in a human hepatoblastoma cell line (HuH-6 clone 5).

Akita M, Tanaka K, Murai N, Matsumoto S, Fujita K, Takaki T, Nishiyama H - Microsc. Res. Tech. (2013)

Localization of CD133 using phase contrast microscopy and ASEM. a: Phase contrast micrograph showing a lamellipodium (arrowhead) and filopodia (arrows). Scale bar, 40 µm. b, c: Localization of CD133 by using ASEM. Cell-surface CD133 was labeled with nanogold, followed by gold enhancement. The clustered particles form a white spot approximately 50 nm in size, mainly in the lamellipodium and filopodia. Scale bar, 1 µm. b: Enlarged ASEM image of the lamellipodium indicated by an arrowhead in Figure 4a. c: Enlarged ASEM image of the filopodia indicated by arrows in Figure 4a.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3842112&req=5

fig04: Localization of CD133 using phase contrast microscopy and ASEM. a: Phase contrast micrograph showing a lamellipodium (arrowhead) and filopodia (arrows). Scale bar, 40 µm. b, c: Localization of CD133 by using ASEM. Cell-surface CD133 was labeled with nanogold, followed by gold enhancement. The clustered particles form a white spot approximately 50 nm in size, mainly in the lamellipodium and filopodia. Scale bar, 1 µm. b: Enlarged ASEM image of the lamellipodium indicated by an arrowhead in Figure 4a. c: Enlarged ASEM image of the filopodia indicated by arrows in Figure 4a.
Mentions: Under a light microscope, strong positive results were seen for CD133 at the cell periphery (Fig. 1), where it co-localized with F-actin (Figs. 2a–2c). Preliminary examination using ASEM also showed that CD133 was localized in membrane ruffles, which occur at the peripheral regions of the cell (Fig. 3). The ASEM enables direct observation of subcellular structures, unlike phase contrast microscopy. Lamellipodia, characteristic features at the leading edge of the cell, were observed. Filopodia were observed as slender cytoplasmic projections that extended beyond the leading edge of lamellipodia. We labeled cell-surface CD133 with nanogold, followed by gold enhancement. The clustered particles formed a white spot, up to 50 nm in size, mainly in the membrane ruffles formed by lamellipodia and filopodia (Fig. 4). Counterstaining with phosphotungstic acid revealed the details of the cells, and ASEM detected details of the ventral cell surface (Fig. 5). When the Aclar® plastic film was set upside down, the dorsal surface of the cell could be observed. CD133 was mainly localized in microvillus-like plasma membrane protrusions (microspikes) on the dorsal surface of the cells Figs.(6 and 7).

Bottom Line: An antibody against CD133 decreased cell migration.Methyl-β-cyclodextrin treatment decreased cell adhesion as well as lamellipodium and filopodium formation.The results suggest that CD133 membrane localization plays a role in tumor cell adhesion and migration.

View Article: PubMed Central - PubMed

Affiliation: Division of Morphological Science, Biomedical Research Center, Saitama Medical University, Iruma-gun, Saitama, Japan. makita@saitama-med.ac.jp

Show MeSH
Related in: MedlinePlus